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In the third year of the SARS‐CoV‐2 pandemic, little is known about the vaccine‐ and infection‐induced immune response in liver transplant recipients (LTR) and liver cirrhosis patients (LCP). This cross‐sectional study assessed the vaccination coverage, infection rate, and the resulting humoral and cellular SARS‐CoV‐2‐specific immune responses in a cohort of LTR and LCP at the University Medical Center Hamburg‐Eppendorf, Germany between March and May 2023. Clinical and laboratory data from 244 consecutive patients (160 LTR and 84 LCP) were collected via chart review and a patient survey. Immune responses were determined via standard spike(S)‐ and nucleocapsid‐protein serology and a spike‐specific Interferon‐gamma release assay (IGRA). On average, LTR and LCP were vaccinated 3.7 and 3.3 times, respectively and 59.4% of patients received ≥4 vaccinations. Altogether, 68.1% (109/160) of LTR and 70.2% (59/84) of LCP experienced a SARS‐CoV‐2 infection. Most infections occurred during the Omicron wave in 2022 after an average of 3.0 vaccinations. Overall, the hospitalization rate was low (<6%) in both groups. An average of 4.3 antigen contacts by vaccination and/or infection resulted in a seroconversion rate of 98.4%. However, 17.5% (28/160) of LTR and 8.3% (7/84) of LCP demonstrated only low anti‐S titers (<1000 AU/ml), and 24.6% (16/65) of LTR and 20.4% (10/59) of LCP had negative or low IGRA responses. Patients with hybrid immunity (vaccination plus infection) elicited significantly higher anti‐S titers compared with uninfected patients with the same number of spike antigen contacts. A total of 22.2% of patients refused additional booster vaccinations. By spring 2023, high vaccination coverage and infection rate have resulted in a robust, mostly hybrid, humoral and cellular immune response in most LTR and LCP. However, booster vaccinations with vaccines covering new variants seem advisable, especially in patients with low immune responses and risk factors for severe disease.

We demonstrate operation of a micropore based flow cytometer in the radio-frequency range. Apart from simply counting micron sized particles, such as cells, with close to nano-second resolution this counter offers the additional benefit of delivering insight into the intracellular environment. Such non-invasive screening of the cell’s interior based on analysing amplitude and phase of the signal is helpful in characterizing the biological activity of cells. In detail we are using heterodyne mixing to demodulate the temporal impedance changes, which are induced by cells translocating through a micropore embedded in a radio-frequency circuit. This allows us to measure every amplitude and phase modulation induced by a translocation event. Herein, we compare the Jurkat cells (human T lymphocytes) recordings with a control group of polystyrene beads. As the cells are measured on a single cell level, the variations on the measured amplitude and phase signals are used, herein, to sense morphological cell changes in real time.

Brain-on-a-chip (BoC) concepts should consider three-dimensional (3D) scaffolds to mimic the 3D nature of the human brain not accessible by conventional planar cell culturing. Furthermore, the essential key to adequately address drug development for human pathophysiological diseases of the nervous system, such as Parkinson’s or Alzheimer’s, is to employ human induced pluripotent stem cell (iPSC)-derived neurons instead of neurons from animal models. To address both issues, we present electrophysiologically mature human iPSC-derived neurons cultured in BoC applicable microscaffolds prepared by direct laser writing. 3D nanoprinted tailor-made elevated cavities interconnected by freestanding microchannels were used to create defined neuronal networks—as a proof of concept—with two-dimensional topology. The neuronal outgrowth in these nonplanar structures was investigated, among others, in terms of neurite length, size of continuous networks, and branching behavior using z-stacks prepared by confocal microscopy and cross-sectional scanning electron microscopy images prepared by focused ion beam milling. Functionality of the human iPSC-derived neurons was demonstrated with patch clamp measurements in both current- and voltage-clamp mode. Action potentials and spontaneous excitatory postsynaptic currents—fundamental prerequisites for proper network signaling—prove full integrity of these artificial neuronal networks. Considering the network formation occurring within only a few days and the versatile nature of direct laser writing to create even more complex scaffolds for 3D network topologies, we believe that our study offers additional approaches in human disease research to mimic the complex interconnectivity of the human brain in BoC studies.

Sepsis remains one of the most lethal and costly conditions treated in U.S. hospitals, with approximately 50% of cases caused by Gram-negative bacterial infections. Septic shock is induced when lipopolysaccharide (LPS), the main component of Gram-negative outer bacterial membrane, signals through the Toll-like receptor 4 (TLR4) complex. Lethal endotoxemia, a model for septic shock, was induced in WT C57BL6 and TLR4–/–mice by administration of Escherichia coliLPS. WT LPS treated mice showed high morbidity, while PBS treated LPS and treated TLR4–/–mice did not. ANOVA analysis of label-free quantification of longitudinal serum proteome revealed 182 out of 324 proteins in LPS injected WT mice that were significantly changed across four time points (0, 6, 12, and 18 h). No significant changes were identified in the two control groups. From the 182 identified proteins, examples of known sepsis biomarkers were validated by ELISA, which showed similar trends as MS proteomics data. Longitudinal analysis within individual mice produced 3-fold more significantly changed proteins than pair-wise comparison. A subsequent global analysis of WT and TLR4–/–mice identified pathways activated independent of TLR4. These pathways represent possible compensatory mechanisms that allow for control of Gram-negative bacterial infection regardless of host immune status.

Nanostructured substrates such as nanowire arrays form a powerful tool for building next-generation medical devices. So far, human pluripotent stem cell-derived neurons—a revolutionary tool for studying physiological function and modeling neurodegenerative diseases—have not been applied to such innovative substrates, due to the highly demanding nature of stem cell quality control and directed differentiation procedures to generate specialized cell types. Our study closes this gap, by presenting electrophysiologically mature human pluripotent stem cell-derived neurons on a set of nanowires in different patterns and growth densities after only four weeks of maturation—thereof 14 to 16 days on the nanowire arrays. While cell viability is maintained on all nanowire substrates, the settling regime of the cells can be controlled and tuned by the nanowire density from a fakir-like state to a complete nanowire wrapping state. Especially, full electrophysiological integrity of the neurons independent of the settling regime has been revealed by patch clamp experiments showing characteristic action potentials. Based on these results, our protocol has the potential to open new pathways in stem cell research and regenerative medicine utilizing human stem cell-derived neurons on tailor-made nanostructured substrates.

UV irradiation-induced cellular damage is classically associated with apoptosis and is known to result in systemic immunosuppression. How the decision to undergo apoptosis is made following UV is not fully understood. We hypothesize that a central mediator of TLR signaling, MyD88, determines cell fate after UV exposure. Survival after UV of immortalized bone marrow-derived macrophages (BMDM) and ex vivo peritoneal macrophages (PM) from MyD88 germline-deficient mice (MyD88-/-) was significantly higher than wild type (WT) PM. UV-induced apoptosis (DNA laddering) in PM and epidermis of MyD88-/-animals versus WT was decreased. In MyD88-/-PM, decreased cleavage of caspase 3, as well as pro-necroptotic protein, RIP1, and a significant increase in transcription and release of pro-inflammatory TNF-α, suggest that necroptosis, rather than apoptosis, has been initiated. In vivo studies confirm this hypothesis after UV, showing low apoptosis by TUNEL and inflammation in MyD88-/-skin sections. Considering that MyD88 participates in many TLR pathways, BMDM from TLR2-/-, TLR4-/-and WT mice were compared for evidence of UV-induced apoptosis. Only TLR4-/-BMDM and PM had a similar phenotype to MyD88-/-, suggesting that the TLR4–MyD88 axis importantly contributes to cell fate decision. Our study describes a new cellular consequence of MyD88 signaling after UV, and may provide rationale for therapies to mitigate UV-induced immunosuppression.

Manganese chloride (MnCl2) 2.5 is included in the extended metals patch test series to evaluate patients for contact hypersensitivity to this metal salt.

We report the electrical transport properties of a hybrid organic/inorganic diode device consisting of a layer of an organic ferrimagnetic semiconductor V[TCNE]x(x~2, TCNE: tetracyanoethylene; TC~ 400 K, EG ~ 0.5 eV, ~ 10-2S/cm) and a GaAs/AlGaAs p-i-ndiode. Comparison with a control excluding the V[TCNE]x~2reveals that the addition of the V[TCNE]x~2layer shifts the turn-on voltage and ideality of the diode in accordance with bulk V[TCNE]x~2properties. This result has implications for the use of inorganic systems as probes of spin physics in organic and molecular systems.

Measurements were made of jump frequencies of 111In/Cd tracer atoms on the Sn-sublattice in rare-earth tri-stannides having the L12 crystal structure via perturbed angular correlation spectroscopy (PAC). Phases studied were Sn3R (R= La, Ce, Pr, Nd, Sm and Gd). Earlier measurements on isostructural rare-earth tri-indides showed that the dominant diffusion mechanism changed along that series [4]. The dominant mechanism was determined by comparing jump frequencies measured at opposing phase boundary compositions (that is, more In-rich and more In-poor). Jump frequencies were observed to be greater at the In-rich boundary composition in light lanthanide indides and greater at the In-poor boundary composition in heavy-lanthanide indides. These observations were attributed to predominance of diffusion via rare-earth vacancies in the former case and indium vacancies in the latter. Contrary to results for the indides, jump frequencies found in the present work are greater for the Sn-poor boundary compositions of the stannides, signaling that diffusive jumps are controlled by Sn-vacancies. Possible origins of these differences in diffusion mechanisms are discussed.

A new technology is presented to suppress the common‐mode ringing in CRT deflection yokes: Ringing‐Free Wire. The effectiveness is analyzed with our newly developed frequency response measuring method.

Infectious diseases propagated by arthropod vectors, such as tularemia, are commonly initiated via dermal infection of the skin. However, due to the technical difficulties in achieving accurate and reproducible dermal deposition, intradermal models are less commonly used.

ABSTRACTAdjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effects while retaining adequate immunogenicity. Despite the virtually unlimited number of potential sources among bacterial strains, the number of useable compounds within this promising class of adjuvants are few. We have developed bacterial enzymatic combinatorial chemistry (BECC) as a method to generate rationally designed, functionally diverse lipid A. BECC removes endogenous or introduces exogenous lipid A-modifying enzymes to bacteria, effectively reprogramming the lipid A biosynthetic pathway. In this study, BECC is applied within an avirulent strain of Yersinia pestisto develop structurally distinct LOS molecules that elicit differential Toll-like receptor 4 (TLR4) activation. Using reporter cell lines that measure NF-κB activation, BECC-derived molecules were screened for the ability to induce a lower proinflammatory response than Escherichia coliLOS. Their structures exhibit varied, dose-dependent, TLR4-driven NF-κB activation with both human and mouse TLR4 complexes. Additional cytokine secretion screening identified molecules that induce levels of tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) comparable to the levels induced by phosphorylated hexa-acyl disaccharide (PHAD). The lead candidates demonstrated potent immunostimulation in mouse splenocytes, human primary blood mononuclear cells (PBMCs), and human monocyte-derived dendritic cells (DCs). This newly described system allows directed programming of lipid A synthesis and has the potential to generate a diverse array of TLR4 agonist candidates.IMPORTANCEThere is an urgent need to develop effective vaccines against infectious diseases that continue to be major causes of morbidity and mortality worldwide. Making effective vaccines requires selecting an adjuvant to strengthen an appropriate and protective immune response. This work describes a practical method, bacterial enzymatic combinatorial chemistry (BECC), for generating functionally diverse molecules for adjuvant use. These molecules were analyzed in cell culture for their ability to initiate immune stimulatory activity. Several of the assays described herein show promising in vitrocytokine production and costimulatory molecule expression results, suggesting that the BECC molecules may be useful in future vaccine preparations.

A collection of 1571 Pisum introductions was screened for resistance to Bruchus pisorum (L.) in field trials by using percentages of infested pods and seeds as the primary evaluation criteria. These criteria permitted excellent differentiation of pea weevil resistant and susceptible pea cultivars and introductions. Entries with 4% or less infested seeds in 1972 and 1974 field trials were plant introduction (P.I.) nos. 196027, 244241, 244254, 244263, 263026, and 280612. Nonpreference for oviposition was judged the major mechanism contributing to resistance. Peduncle lengths of resistant introductions were significantly shorter than those of susceptible cultivars and were highly correlated with lower percentages of infested pods and seeds.